A minimised hammerhead ribozyme with activity against interleukin-2 in human cells

A "minizyme" is a smaller version of the hammerhead ribozyme, in which stem-loop II has been replaced by a short linker. Here, we have synthesised a DNA-containing minizyme and a ribozyme, which are designed to cut within a 15-nucleotide sequence in human interleukin-2 mRNA, and have tested for their activity in vitro and in cells. In vitro at 37 degrees C, a minizyme with linker of sequence d(GTTTT) cleaves a 15-ribonucleotide synthetic substrate 5-fold slower than does the full-sized ribozyme. In human cells, the minizyme inhibits the production of interleukin-2 protein to a similar extent as does the ribozyme. Also, the minizyme and the ribozyme are more effective in cells than any of three controls: an inactive minizyme, a 15-nucleotide antisense DNA, or DNA of random sequence. The positive effect observed in cells indicates that minizymes may be useful as pharmaceuticals.